| 摘要: |
| [目的]从猪血清7型胸膜肺炎放线杆菌中提取纯化尿素酶,并对其生物特性和理化特性进行研究.[方法]超声破碎放线杆菌粗提尿素酶,再采用硫酸铵盐析和Sephadex G-200凝胶过滤层析对该酶进行纯化,分析其活性及理化特性.[结果]最适于猪胸膜肺炎放线杆菌尿素酶保持活性的pH为8.5,温度为45 ℃.在该条件下其比酶活约为96.253 U/mg;SDS-PAGE电泳显示,胸膜肺炎放线杆菌尿毒酶含分子质量为11,25和60 ku的亚单位.[结论]与传统方法相比,超声破壁和Sephadex G-200凝胶过滤层析纯化法简单快速,重复性好,测定尿素酶的特性稳定,能满足实验室检测的要求. |
| 关键词: 猪传染性胸膜肺炎 胸膜肺炎放线杆菌 尿素酶 超声破壁 酶活性 理化特性 |
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| 基金项目:山东农业大学科技创新基金项目（23414） |
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| Purification and characterization of urease from Actinobacillus pleuropneumoniae |
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| Abstract: |
| 【Objective】 Urease from Actinobacillus pleuropneumoniae serotype 7 was purified to study its physical and biochemical proprties.【Method】 Urease from Actinobacillus pleuropneumoniae serotype 7 was extracted,then (NH4)2SO4 fractionation and Sephadex G-200 gel filtration were used to extract and purify urease.【Result】 The optimum pH and temperature for the urease to keep the activities were 8.5 and 45 ℃.The specific activity of urease under this situation was 96.253 U/mg.The urease had three subunits,and the molecular weight for each was about 11,25 and 60 ku by SDS-PAGE.【Conclusion】 Using ultrasonication and Sephadex G-200 gel filtration to extract urease was a simple and quick meature,the results of which were stable and could reach the requirement of experiment properties. |
| Key words: Porcine contagious pleuropneumonia Actinobacillus pleuropneumoniae urease ultrasonication activity of urease physical and biochemical properties |
